392 replies to this topic

[Oakman]

For people who didn't bother so far reading up about "NAD-boosters" (like myself) - this new episode (no guests) of R.Patrick gives a good general overview:

 

https://www.foundmyf...odes/nad-nr-nmn

 

 

takeaway from animal studies:

 

1. at low oral dose, neither NMN nor NR are bioavailable beyond the liver

 

2. at high oral dose, there is some very limited bioavailability; but it's not generally making it out of the liver

 

More critical: at either dose, all/most of the NMN/NR that is not used up by liver cells, is metobalised into nicotimanide. The latter is part of feedback-loop that limits the endogenous production of NAD+. So you're externally upping nicotimanide and hence NAD+, but at the same time lowering endogenous production of it - maintaining a "natural" steady-state.

 

 

3. by injecting a high dose of NR (avoiding the first pass effect of the liver), kidneys and muscles are able to directly form NAD+. But it's still not able to go beyond the blood-brain-barrier.

 

 

 

I am curious: why is anyone at all taking oral NAD-boosters? Even ignoring data of detrimental effects on possibly cancer, Sirt-1 etc. If anyone is serious about NAD-boosters: why aren't you all buying needles and inject yourself a high dose every day to have any effect at all? Oral intake seems to be a complete waste of money. What am I missing?

 

Well, the fact that no one, nobody, none of us, really knows if there is benefit or not. Many THINK there is, and so with free choice many think the cost in worth being in the game, rather than kibitzing from the sidelines. Reality rocks!

[Smith]

For people who didn't bother so far reading up about "NAD-boosters" (like myself) - this new episode (no guests) of R.Patrick gives a good general overview:

 

https://www.foundmyf...odes/nad-nr-nmn

 

 

takeaway from animal studies:

 

1. at low oral dose, neither NMN nor NR are bioavailable beyond the liver

 

2. at high oral dose, there is some very limited bioavailability; but it's not generally making it out of the liver

 

More critical: at either dose, all/most of the NMN/NR that is not used up by liver cells, is metobalised into nicotimanide. The latter is part of feedback-loop that limits the endogenous production of NAD+. So you're externally upping nicotimanide and hence NAD+, but at the same time lowering endogenous production of it - maintaining a "natural" steady-state.

 

 

3. by injecting a high dose of NR (avoiding the first pass effect of the liver), kidneys and muscles are able to directly form NAD+. But it's still not able to go beyond the blood-brain-barrier.

 

 

 

I am curious: why is anyone at all taking oral NAD-boosters? Even ignoring data of detrimental effects on possibly cancer, Sirt-1 etc. If anyone is serious about NAD-boosters: why aren't you all buying needles and inject yourself a high dose every day to have any effect at all? Oral intake seems to be a complete waste of money. What am I missing?

 

Based upon all the recent evidence, oral supplementation of NR/NMN is really no different than oral supplementation of NAM... that is unless you are willing to take large amounts of NR/NAM to overwhelm the liver's ability to metabolize it. 

[JamesPaul]

Can NR or NMN be absorbed sub-lingually and if so, can they be converted into NAD+ without being limited by the body's rate-limiting mechanisms, at least on the first pass through the circulatory system?

[Linux]

Can NR or NMN be absorbed sub-lingually and if so, can they be converted into NAD+ without being limited by the body's rate-limiting mechanisms, at least on the first pass through the circulatory system?

 

There´s no scientific evidence that NR or NMN can be absorbed sublingually. None of the human (or animal) studies on either compound used sublingual administration.

 

There are studies on oral administration of NR and NMN  (in humans and animals) and injected (animals) but not using the sublingual route. Lack of scientific evidence doesn´t stop companies from claiming whatever they want in their promotional material though. 

 

Sublingual NR/NMN or time-release mechanisms are just ways to differentiate your product from others on the market. Non evidence-based gimmicks.

 

 

Edited by Linux, 17 November 2019 - 12:05 PM.

[able]

There´s no scientific evidence that NR or NMN can be absorbed sublingually. None of the human (or animal) studies on either compound used sublingual administration.

 

There are studies on oral administration of NR and NMN  (in humans and animals) and injected (animals) but not using the sublingual route. Lack of scientific evidence doesn´t stop companies from claiming whatever they want in their promotional material though. 

 

Sublingual NR/NMN or time-release mechanisms are just ways to differentiate your product from others on the market. Non evidence-based gimmicks.

 

 

You're right, there is certainly no studies yet published on sublingual NMN.

 

But I would argue something is not a gimmick if it actually works.

 

Just because there are not yet any clinical studies, does not prove it is ineffective.

 

It seems that the great majority of people that have tried it can feel the effect within minutes, which is not possible unless some  is aborbed.  

 

I have posted many times on this board that I get an immediate measurable response of slightly elevated heart rate minutes after taking multiple dosages of the powder.

 

I would agree that we have no clue what the effect is of NMN absorbed sublingually, but some is definately absorbed that way.

 

Looking forward to studies on this.

[Linux]

You're right, there is certainly no studies yet published on sublingual NMN.

 

It seems that the great majority of people that have tried it can feel the effect within minutes, which is not possible unless some  is aborbed.  

 

I have posted many times on this board that I get an immediate measurable response of slightly elevated heart rate minutes after taking multiple dosages of the powder.

 

The effect people feel "within minutes" of taking supplements is indeed real.

 

It is called the placebo effect.

 

Unless your supplement is tainted with sildenafil, amphetamine, nicotinic acid or any other fast-acting drugs you don´t "feel" vitamins working within minutes, if at all.

 

The recent NMN safety-trial revealed NO changes in heart rate. I´m quoting:

 

"The single oral administrations of NMN did not cause any significant clinical symptoms or changes in heart rate, blood pressure, oxygen saturation, and body temperature."

 

 

What you may have experienced, besides the placebo effect, may be a small amount of nicotinic acid in your powder. A small flush can raise the pulse in some individuals. But it is probably just excitement over taking this expensive longevity-powder that raises your pulse.

Edited by Linux, 17 November 2019 - 04:51 PM.

[able]

The effect people feel "within minutes" of taking supplements is indeed real.

 

It is called the placebo effect.

 

Unless your supplement is tainted with sildenafil, amphetamine, nicotinic acid or any other fast-acting drugs you don´t "feel" vitamins working within minutes, if at all.

 

The recent NMN safety-trial revealed NO changes in heart rate. I´m quoting:

 

 

What you may have experienced, besides the placebo effect, may be a small amount of nicotinic acid in your powder. A small flush can raise the pulse in some individuals. But it is probably just excitement over taking this expensive longevity-powder that raises your pulse.

 

 

Yes, placebo effect is strong.  

 

More so in longer term effects like "my knees feel so much better the last 2 weeks since I have been taking curcumin"

 

Not so relevant when noting a strong cup of coffee really wakes you up.

 

Sublingual NMN is more the latter - very similar to a cup of coffee but stronger.

 

Of course the human study with NMN found no effect on heart rate, as they used capsules, which go thru stomach  and liver first.

 

As for being spiked with niacin - both the ABN and Prohealth brands give the same effect, and both post test results from 3rd party labs. So unless both are doing it, and the testing companies are complicit, that doesn't seem possible.

 

Not to mention that the effect is much faster than possible from taking any capsule - niacin or whatever.

Edited by able, 17 November 2019 - 05:52 PM.

[Andey]

Yes, placebo effect is strong.  

 

More so in longer term effects like "my knees feel so much better the last 2 weeks since I have been taking curcumin"

 

Not so relevant when noting a strong cup of coffee really wakes you up.

 

Sublingual NMN is more the latter - very similar to a cup of coffee but stronger.

 

Of course the human study with NMN found no effect on heart rate, as they used capsules, which go thru stomach  and liver first.

 

As for being spiked with niacin - both the ABN and Prohealth brands give the same effect, and both post test results from 3rd party labs. So unless both are doing it, and the testing companies are complicit, that doesn't seem possible.

 

Not to mention that the effect is much faster than possible from taking any capsule - niacin or whatever.

 

TBH I dont feel anything distinctive from ABN sublinguals, both NAD and NMN

But I feel ABNs NAD nasal spray

There is also a difference between those NAD, NMN and NAM + ribose. From my recollection NAM + ribose dont affect my sleep latency, but sublinguals are messing it  up.

Its all n=1 though

[MikeDC]

Using pharmacological inhibitors of plasma membrane transporters, we also showed that extracellular cleavage of NAD+ and NMN to NR is a prerequisite for using these nucleotides to maintain intracellular NAD contents.

https://www.ncbi.nlm...1/?i=1&from=nmn

[Linux]

Using pharmacological inhibitors of plasma membrane transporters, we also showed that extracellular cleavage of NAD+ and NMN to NR is a prerequisite for using these nucleotides to maintain intracellular NAD contents.

https://www.ncbi.nlm...1/?i=1&from=nmn

 

The last sentence about NR both hydrolyzing and being intracellularly converted into nicotinamide got cut out by mistake Mike DC. I fixed it for you by supplying the whole quote:

 

"Using pharmacological inhibitors of plasma membrane transporters, we also showed that extracellular cleavage of NAD+ and NMN to NR is a prerequisite for using these nucleotides to maintain intracellular NAD contents. We also present evidence that, besides spontaneous hydrolysis, NR is intensively metabolized in cell culture by intracellular conversion to Nam."

 

I don´t think cell culture studies should be used as proof for anything really. We are a whole system, not compartments of individual cells in a petri dish.

Edited by Linux, 06 December 2019 - 05:22 AM.

[MikeDC]

The last sentence about NR both hydrolyzing and being intracellularly converted into nicotinamide got cut out by mistake Mike DC. I fixed it for you by supplying the whole quote:

"Using pharmacological inhibitors of plasma membrane transporters, we also showed that extracellular cleavage of NAD+ and NMN to NR is a prerequisite for using these nucleotides to maintain intracellular NAD contents. We also present evidence that, besides spontaneous hydrolysis, NR is intensively metabolized in cell culture by intracellular conversion to Nam."

I don´t think cell culture studies should be used as proof for anything really. We are a whole system, not compartments of individual cells in a petri dish.

Since NMN is required to covert to NR in order to enter cells, NMN will face the same intracellular inversion issue.

[LawrenceW]

Since NMN is required to covert to NR in order to enter cells, NMN will face the same intracellular inversion issue.

 

MikeDC

 

It is time for you to stop regurgitating Brenner's debunked anti-NMN talking points.

 

https://www.nature.c...2255-018-0009-4

 

"We have previously demonstrated that the transport of NMN from the gut to the circulation and then to tissues occurred within 10 min (ref. 15). However, the mechanism that mediates such minute-order transport of NMN has so far remained unknown. In this study, we demonstrate that the Slc12a8 gene encodes a novel NMN transporter in mammals. The mRNA expression of the Slc12a8 gene is upregulated in response to NAD+ decline, allowing cells to meet to an urgent demand for NAD+ biosynthesis. The Slc12a8 NMN transporter is specific to NMN"

[MikeDC]

MikeDC

It is time for you to stop regurgitating Brenner's debunked anti-NMN talking points.

https://www.nature.c...2255-018-0009-4

"We have previously demonstrated that the transport of NMN from the gut to the circulation and then to tissues occurred within 10 min (ref. 15). However, the mechanism that mediates such minute-order transport of NMN has so far remained unknown. In this study, we demonstrate that the Slc12a8 gene encodes a novel NMN transporter in mammals. The mRNA expression of the Slc12a8 gene is upregulated in response to NAD+ decline, allowing cells to meet to an urgent demand for NAD+ biosynthesis. The Slc12a8 NMN transporter is specific to NMN"

Only this research group claims there is a NMN transporter and even they admit the amount of NMN transported is very small. In their first paper they only observed the initial absorption and the total NMN accounted for is only 5%. The rest is absorbed later and they did not observe. Liu’s Paper also show a little amount of NR was absorbed in the initial period. So this is unlikely due to an NMN transporter. Most like small absorption in the stomach.
Brenner criticized the second study as making things up with noise.
All other papers from different research groups conclude that NMN is required to convert to NR before getting into cells.

[Harkijn]

I really hope for the NMN transporter to exist but we really need some duplication, preferably by another research group. I find it curious how little scientific attention NMN draws.  AFAIK we only recently saw a first human safety study, concluding safety after... a one time dose.

[MikeDC]

Be careful what you wish for! If NMN has a decent transporter across cell membranes, NMN users would have already got neurological diseases because high levels of NMN activates SARM1 which depletes NAD+ inside cells.
https://www.ncbi.nlm...les/PMC6531917/

Edited by MikeDC, 06 December 2019 - 10:03 PM.

[Linux]

MikeDC,

So by your logic both NR and NMN is neurotoxic? (there's no data supporting that either metabolite is neurotoxic on its own).

Because all NR must get phosphorylated into NMN within mammalian cells before turning into NAD.

Edited by Linux, 06 December 2019 - 10:42 PM.

[MikeDC]

MikeDC,

So by your logic both NR and NMN is neurotoxic? (there's no data supporting that either metabolite is neurotoxic on its own).

Because all NR must get phosphorylated into NMN within mammalian cells before turning into NAD.

Only high levels of NMN is neurotoxic. Oral NR can’t generate high level NMN inside cells because the conversion of NR to NMN is limited by NRK1.

[Linux]

Dr Brenner and colleagues showed that NMN administration IN ITSELF was not neurotoxic and caused no degeneration:

"Collectively, our data indicate that maintaining NAD+ is not sufficient to protect DRG neurons from vincristine-induced axon degeneration, and elevating NMN, by itself, is not sufficient to cause degeneration."

https://pubmed.ncbi....e=10&from_pos=1

[able]

Using pharmacological inhibitors of plasma membrane transporters, we also showed that extracellular cleavage of NAD+ and NMN to NR is a prerequisite for using these nucleotides to maintain intracellular NAD contents.

https://www.ncbi.nlm...1/?i=1&from=nmn

 

 

 

Keep in mind, they are only speaking of this one particular type of cell.

 

And, even in this cell they found there was no obstacle to entering cells, regardless the route taken.

 

They disabled the salvage pathway with fk866 which stops NAD+ creation, and added different NAD+ metabolites.

 

They found NMN and NAD did indeed rescue NAD+ inside the cells.  See chart below.

 

In fact NMN was more effective than NR or any other metabolite.

 

More proof of what I have long suspected - any theoretical advantage NR might have at crossing the cell membrane in some cell types is insignificant.

 

Also note that advantage is totally meaningless if NR is not readily available in the bloodstream AS NR, which is what all studies have found so far.  

 

The authors in this study were surprised at how unstable NR is and how quickly NR was hydrolyzed by water or plasma to NAM.  

 

Which explains why NR is either not found at all, or only at trace levels in the blood.

 

 

 

 

 

 

Figure 2. Extracellular NAD+ intermediates support NAD+ generation in HEK293 cells. HEK293 cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM) containing Nam, supplemented with 10% fetal bovine serum (FBS). To inhibit NAD+ synthesis from Nam, cells were treated with FK866. Cells were also treated with NAD+ or its derivatives as indicated. Metabolic activity was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay 48 h after the treatment. Metabolic activity of untreated cells (control) was taken as 100%. Data are presented as mean ± S.D (n = 3). Statistical analysis of differences between the groups was carried out by one-way ANOVA with post hoc comparisons using Tukey test. * indicates statistical difference at p < 0.001.

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[MikeDC]

Dr Brenner and colleagues showed that NMN administration IN ITSELF was not neurotoxic and caused no degeneration:

"Collectively, our data indicate that maintaining NAD+ is not sufficient to protect DRG neurons from vincristine-induced axon degeneration, and elevating NMN, by itself, is not sufficient to cause degeneration."

https://pubmed.ncbi....e=10&from_pos=1

The new study says elevating NMN high enough is sufficient to activate SARM1 to degrade NAD+

[MikeDC]

Keep in mind, they are only speaking of this one particular type of cell.

And, even in this cell they found there was no obstacle to entering cells, regardless the route taken.

They disabled the salvage pathway with fk866 which stops NAD+ creation, and added different NAD+ metabolites.

They found NMN and NAD did indeed rescue NAD+ inside the cells. See chart below.

In fact NMN was more effective than NR or any other metabolite.

More proof of what I have long suspected - any theoretical advantage NR might have at crossing the cell membrane in some cell types is insignificant.

Also note that advantage is totally meaningless if NR is not readily available in the bloodstream AS NR, which is what all studies have found so far.

The authors in this study were surprised at how unstable NR is and how quickly NR was hydrolyzed by water or plasma to NAM.

Which explains why NR is either not found at all, or only at trace levels in the blood.






Figure 2. Extracellular NAD+ intermediates support NAD+ generation in HEK293 cells. HEK293 cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM) containing Nam, supplemented with 10% fetal bovine serum (FBS). To inhibit NAD+ synthesis from Nam, cells were treated with FK866. Cells were also treated with NAD+ or its derivatives as indicated. Metabolic activity was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay 48 h after the treatment. Metabolic activity of untreated cells (control) was taken as 100%. Data are presented as mean ± S.D (n = 3). Statistical analysis of differences between the groups was carried out by one-way ANOVA with post hoc comparisons using Tukey test. * indicates statistical difference at p < 0.001.

No, this study says NMN can’t enter cells. But their own modified NMN derivative can.

[Oakman]

"Collectively, our data indicate that maintaining NAD+ is not sufficient to protect DRG neurons from vincristine-induced axon degeneration, and elevating NMN, by itself, is not sufficient to cause degeneration."

 

The study was about "vincristine", a chemo drug, and its effects on dorsal root ganglia. What does that have to do with normal metabolism...not much. Plus NMN alone does not cause degradation. End result...this study, like many specific case studies on disease models, provides insight only to the case at hand.

[able]

No, this study says NMN can’t enter cells. But their own modified NMN derivative can.

 

I see no reference to an NMN derivative.  Can you please point to that?

 

They simply added different NAD+ metabolites to the serum and observed they were all able to rescue NAD+ metabolism inside the cells, with standard NMN being more effective than any other.

 

The route they take to enter cells does not matter at all.  NMN is more effective than all other NAD+ metabolites.

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Edited by able, 07 December 2019 - 04:24 PM.

[MikeDC]

Read the title of the paper.

https://www.ncbi.nlm...les/PMC6531917/

Our results show that both CZ-48 and NMN can induce the same conformational changes in SARM1 and activate its enzymatic activities. However, NMN can do so only in cell lysates but not in intact cells (Figures 3A and 3C), indicating it is not permeable to the cells we tested

Edited by MikeDC, 07 December 2019 - 08:22 PM.

[able]

Read the title of the paper.

https://www.ncbi.nlm...les/PMC6531917/

Our results show that both CZ-48 and NMN can induce the same conformational changes in SARM1 and activate its enzymatic activities. However, NMN can do so only in cell lysates but not in intact cells (Figures 3A and 3C), indicating it is not permeable to the cells we tested

 

 

Why are you referencing a different study?

 

You posted about this study:

 

 

Degradation of Extracellular NAD⁺Intermediates in Cultures of Human HEK293 Cells

 

I replied about that study, with quotes and a chart about that study.

 

You said that chart was not valid as they used an NMN derivative.  I said it was plain NMN. Why are you telling me to read the title of a different study?

Edited by able, 07 December 2019 - 08:52 PM.

[MikeDC]

Why are you referencing a different study?

You posted about this study:


Degradation of Extracellular NAD⁺Intermediates in Cultures of Human HEK293 Cells

I replied about that study, with quotes and a chart about that study.

You said that chart was not valid as they used an NMN derivative. I said it was plain NMN. Why are you telling me to read the title of a different study?

I linked the correct study. However when you quoted my post you replaced my link with a different study.

[able]

Ok, lets try this again.

 

Degradation of Extracellular NAD+ Intermediates in Cultures of Human HEK293 Cells

 

THIS study shows NMN and NAD+ must convert to NR to enter the particular cell type used in this one study. Hooray, exactly what Dr Brenner claims is the reason NR is the best precursor. Although many studies have shown NMN and NAD+ do enter some cell types directly, we'll just look at this one.

 

Keep in mind, they are only speaking of this one particular type of cell.

 

And, even in this cell they found there was no obstacle to entering cells, regardless the route taken.

 

There are ecto-enzymes on the outside of cells that convert NAD+ and metabolites.

To test the uptake, they disabled the salvage pathway with fk866 which stops internal NAD+ creation.  Then they added different NAD+ metabolites to plasma outside the cells to see how if they could rescue NAD+ metabolism inside cells with the salvage pathway disabled.

They found NMN and NAD did indeed rescue NAD+ inside the cells. See chart below.

In fact NMN was more effective than NR or any other metabolite.

More proof of what I have long suspected - any theoretical advantage NR might have at crossing the cell membrane in some cell types is insignificant.

Also note that advantage is totally meaningless if NR is not readily available in the bloodstream AS NR, which is what all studies have found so far.  

The authors in this study also note how unstable NR is and how quickly NR was hydrolyzed by water or plasma to NAM.  

 

Likewise, we observed cleavage of NR to Nam in the medium without cells. 

 

Strikingly, in the control sample that contained water instead of FBS, the extent of NR cleavage was similar (Figure 4, right lower panel). 

 

These data indicate that NR efficiently hydrolyses to Nam (and ribose) in aqueous solutions

 

Which explains why NR is either not found at all, or only at trace levels in the blood in all studies so far.





 

Figure 2. Extracellular NAD+ intermediates support NAD+ generation in HEK293 cells. HEK293 cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM) containing Nam, supplemented with 10% fetal bovine serum (FBS). To inhibit NAD+ synthesis from Nam, cells were treated with FK866. Cells were also treated with NAD+ or its derivatives as indicated. Metabolic activity was measured by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay 48 h after the treatment. Metabolic activity of untreated cells (control) was taken as 100%. Data are presented as mean ± S.D (n = 3). Statistical analysis of differences between the groups was carried out by one-way ANOVA with post hoc comparisons using Tukey test. * indicates statistical difference at p < 0.001.

 

 

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Edited by able, 07 December 2019 - 11:17 PM.

[MikeDC]

First you admit that the paper says NMN needs to convert to NR before entering cells. Then you say NMN can enter cells with no problem. Yes, NMN can increase NAD+ inside cells by changing to NR first. You cannot reason that NMN has no problem entering cells because it increases intracellular NAD+.

[able]

First you admit that the paper says NMN needs to convert to NR before entering cells. Then you say NMN can enter cells with no problem. Yes, NMN can increase NAD+ inside cells by changing to NR first. You cannot reason that NMN has no problem entering cells because it increases intracellular NAD+.

 

 

Yes, clearly this study agrees with others that in SOME cells, NMN and NAD must first convert to NR.

 

What THIS STUDY also shows, is that it isn't an obstacle and NMN is still more effective at restoring NAD+ metabolism inside the cells.

 

They disabled internal NAD+ salvage, so the cells could only get NAD+ from external sources.

 

They added the various NAD+ metabolites OUTSIDE the cells.

 

They found that NMN added outside the cells was more effective than NR and all other metabolites at restoring NAD+ INSIDE the cells.  See chart below.

 

How could it be more effective than NR?  They note NMN is fairly stable, while NR is easily degraded to NAM in plasma or water.  Just like Dr Sinclair has been saying for a few years now.

 

 

 

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Edited by able, 08 December 2019 - 12:20 AM.

[MikeDC]

It is really not new that NMN can increase NAD+ inside cells by converting to NR first. Yes, in this specific cell study, NMN seems to be slightly productive on equal molar basis. Remember NMN is 31% heavier than NR. If you normalize to equal weight basis, NR will be more effective.